Use of Nucleoporin-Conjugated Beads to Study the Nuclear Pore Complex Assembly on the Nuclear Membrane.

Reassembly of the nuclear pore complex (NPC) at the end of mitosis is a key event for establishing a functional nuclear envelope. However, the role of each NPC component (Nup) in NPC reassembly remained unclear. Here, we describe a method using artificial beads conjugated with an anti-GFP antibody. In this method, such beads are introduced into living cells expressing the Nup of interest fused with GFP to tether the specific Nup around the beads. By examining the Nup-conjugated beads in mitotic telophase cells, it is possible to investigate the role of the specific Nup of interest in NPC formation on the nuclear membrane.